γE-crystallin CRISPR/Cas9 KO Plasmid (m): sc-419832
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- γE-crystallin CRISPR/Cas9 Knockout (KO) Plasmid (m) is a pool of plasmids, each encoding Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed for maximum knockout efficiency using sequences derived from the GeCKO v2 library
- gRNA sequences direct Cas9 to induce site-specific double-strand breaks (DSBs) in the γE-crystallin genomic locus, resulting in gene knockout through non-homologous end joining (NHEJ)
- The puromycin resistance and RFP genes are flanked by LoxP sites, enabling removal of selection markers via Cre recombinase (Cre Vector: sc-418923) after establishing stable knockout cell lines
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
γE-crystallin CRISPR/Cas9 KO Plasmid (m) | sc-419832 | 20 µg | $397.00 |
Overview
Cryge encodes γE-crystallin, a lens fiber cell structural protein within the β/γ-crystallin superfamily that contributes to the exceptionally high refractive index and long-term transparency of the ocular lens. γE-crystallin participates in the tightly regulated process of lens fiber cell differentiation and maturation, where crystallin packing, protein stability, and resistance to aggregation are central to proteostasis. Disruption of crystallin homeostasis can promote light-scattering aggregates and compromise lens biomechanics, linking crystallin gene perturbation to cataract-associated phenotypes and related lens opacification mechanisms. In mouse models, Cryge is studied to understand crystallin network organization, post-translational modifications, and stress responses that influence lens clarity over time.
γE-crystallin CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Cryge gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Cryge together with the Streptococcus pyogenes Cas9 nuclease. The plasmids also encode GFP, allowing fluorescent identification and enrichment of successfully transfected cells by fluorescence microscopy or flow cytometry.
The multi-guide design increases the likelihood of generating insertions or deletions (indels) that disrupt the Cryge open reading frame following Cas9-mediated double-strand break formation. DNA breaks introduced by the CRISPR/Cas9 system are repaired through endogenous non-homologous end joining (NHEJ) pathways, frequently resulting in frameshift mutations that abolish γE-crystallin protein expression.
This CRISPR knockout system enables efficient generation of Cryge-deficient cell models for investigation of γE-crystallin signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Cryge exon(s) critical for γE-crystallin function
- Co-expression of SpCas9 and sgRNA from a single plasmid for simplified delivery
- GFP reporter for identification of transfected cells
- Pool of plasmids targeting multiple Cryge genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by γE-crystallin CRISPR/Cas9 KO Plasmid (m) and γE-crystallin CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Cryge locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by γE-crystallin HDR Plasmid (m) and γE-crystallin HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Cryge homology arms to support homology-directed repair at defined Cryge target sites corresponding to the CRISPR/Cas9 KO designs. HDR donor availability may vary. See Related Products for availability.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.